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1.
Comparison of nine commercial immunoassays for the detection of rotavirus in fecal specimens.
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One hundred fecal specimens obtained from patients with acute gastroenteritis were tested for rotavirus with nine commercial immunoassays to evaluate the sensitivity, specificity, predictive value, and diagnostic accuracy of these assays. Kits evaluated included two monoclonal antibody-based enzyme immunoassays (EIAs) (Rotaclone and Pathfinder Rotavirus), three polyclonal antibody-based EIAs (Rotavirus Immunoassay, Rotazyme II, and Wellcozyme Rotavirus), and four latex agglutination assays (Rotastat, Virogen Rotatest, Meritec-Rotavirus, and The Wellcome Latex Test). Thirty-eight of the 100 specimens were found to contain rotavirus by a reference microplate EIA. The accuracy of the reference assay was determined by RNA electrophoresis and a blocking assay on discordant specimens. The two monoclonal antibody EIAs had superior sensitivities (100%) and identified two positive specimens which were negative by the reference method but positive by the blocking assay. Among the polyclonal EIAs, all had sensitivities of greater than 90%, but specificities were variable; Rotazyme II, with a specificity of 50%, showed considerable discrepancy from other polyclonal EIAs. The latex tests had sensitivities ranging from 70 to 90% and specificities of 80 to 100%. Latex agglutination tests were more rapid than EIAs and did not require expensive equipment. The final choice of assay system will depend on the cost, speed, and accuracy requirements of the clinical laboratory. 相似文献
2.
J L Cromien C A Himmelreich R I Glass G A Storch 《Journal of clinical microbiology》1987,25(12):2359-2362
We evaluated a new commercial enzyme immunoassay (EIA) for rotavirus (Rotavirus EIA; International Diagnostic Laboratories, Chesterfield, Mo.). A total of 161 consecutive stool samples (including 18 from infants less than 30 days old) submitted to the diagnostic laboratory at Children's Hospital, Washington University Medical Center, St. Louis, Mo., for rotavirus detection were tested by Rotavirus EIA and by Rotazyme II (Abbott Laboratories, North Chicago, III.) according to the instructions of the manufacturer. In addition, 16 samples from infants less than 30 days old without diarrhea were tested by both assays. Samples showing discrepant results after repeat testing were examined by electron microscopy. Nine samples yielding discrepant results were also tested by using a reference EIA directly on the specimen and on culture supernatants from two passages in MA 104 cells. Rotavirus EIA and Rotazyme II yielded concordant results for 85% of the samples. All of the 26 discrepant samples tested negative by Rotavirus EIA and positive (15 samples) or equivocal (11 samples) by Rotazyme II. These samples included 11 from symptomatic infants more than 30 days old, 2 from symptomatic infants less than 30 days old (neonates), and 2 from neonates without diarrhea. Rotavirus was not detected in any of the 24 that were examined by electron microscopy or in any of the 9 that were tested by the reference EIA. The sensitivity, specificity, positive predictive value, and negative predictive value were 100% for Rotazyme EIA and 100, 90, 70, and 100%, respectively, for Rotazyme II. Rotavirus EIA was comparable to Rotazyme II in ease of performance. We conclude that Rotavirus EIA is equally sensitive and more specific than Rotazyme II for detecting rotavirus. Rotavirus EIA is a practical and accurate rotavirus assay for use in clinical laboratories. 相似文献
3.
Detection of rotavirus in stool specimens with monoclonal and polyclonal antibody-based assay systems. 总被引:3,自引:9,他引:3
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Accurate diagnosis of rotavirus is important in both clinical and research situations. A total of 100 stool specimens from children with diarrhea were tested for rotavirus by electron microscopy. These specimens were then coded and tested for rotavirus by four procedures: a monoclonal antibody-based enzyme immunoassay (EIA) (Pathfinder; Kallestad Laboratories, Inc., Austin, Tex.), two polyclonal antibody-based EIAs (Rotazyme II; Abbott Laboratories, North Chicago, Ill.; and an EIA performed with reagents from the National Institutes of Health, Bethesda, Md. [NIH reagent EIA]), and a latex agglutination (LA) assay (Rotalex; Medical Technology Corp., Somerset, N.J.). The sensitivity of the monoclonal antibody EIA (95%) was superior to those of the polyclonal antibody EIAs (73% for Rotazyme II and 57% for the NIH reagent EIA) and the LA assay (61%). The specificity of the LA assay (98%) was slightly better than those of the other systems (88 to 96%). The positive and negative predictive values of the monoclonal antibody EIA (93 and 96%, respectively) were better than those of Rotazyme II (82 and 80%, respectively), the LA assay (96 and 76%, respectively), and the NIH reagent EIA (93 and 74%, respectively). The visual readings of the monoclonal antibody EIA correlated better with the spectrophotometric optical density readings than did the visual readings of the polyclonal antibody EIAs; however, the agreement of both with electron microscopy results was poor when 1+ or plus-minus readings were observed. The monoclonal antibody EIA is more sensitive and predictive than other rotavirus detection systems and second only to the LA assay in specificity in detecting rotavirus in stool specimens. 相似文献
4.
A latex agglutination (LA) test was evaluated for the detection of human rotaviruses in stool specimens. Both antiserum and immunoglobulin G (IgG)-sensitized latex particles were used, with IgG-coated beads being more sensitive for human rotavirus antigen detection. Latex beads sensitized with anti-simian-SA-11 IgG were stable for at least 8 months when stored at 4 degrees C. The sensitivity of the test was compared with that of the Rotazyme (Abbott Laboratories, Diagnostics Div., North Chicago, Ill.) test. The least number of particles detected was 9.0 X 10(5) particles by the LA test versus 4.5 X 10(5) particles by the Rotazyme test. When 10 stool specimens were serially diluted for antigen endpoint determinations, the geometric mean titer by the LA test was 592 versus 1,280 by the Rotazyme test. Forty-three stool samples positive by the Rotazyme test were all positive by the LA test, and no false negative results were detected. Unconfirmed false positive reactions ranged between 8 and 24%. The LA test for rotavirus antigen detection is direct, easy to perform, sensitive, quick, and may have application for use in diagnostic laboratories, emergency rooms, and physician's offices. 相似文献
5.
Polyacrylamide gel electrophoresis of RNA compared with polyclonal- and monoclonal-antibody-based enzyme immunoassays for rotavirus.
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D L Pacini M T Brady C T Budde M J Connell V V Hamparian J H Hughes 《Journal of clinical microbiology》1988,26(2):194-197
Polyacrylamide gel electrophoresis (PAGE) of rotaviral RNA, a sensitive and highly specific test for detecting rotavirus in stool, was compared with two commercially available enzyme immunoassays (EIAs), monoclonal (Pathfinder) and polyclonal (Rotazyme II). Stool samples from 204 children with nosocomial diarrhea were tested for rotavirus by both EIAs and by PAGE of RNA extracted from raw stools or 10% stool suspensions. Samples which tested positive by either EIA but were negative by PAGE were subjected to blocking EIA with rabbit or goat anti-SA11. Rotavirus was detected by PAGE and Pathfinder in 62 stools, but only 47 of these were positive by Rotazyme II. Blocking assays of EIA-positive, PAGE-negative samples suggested the presence of rotavirus in four additional stools. Sensitivity and specificity measured against PAGE and blocking assays were: Pathfinder, 0.985 and 0.934; and Rotazyme II, 0.731 and 0.927, respectively. False-positive rates were 0.134 for Pathfinder and 0.149 for Rotazyme II. The specificity and rate of false-positive results of Pathfinder were improved by using an adjusted optical density cutoff 4.36 times greater than that recommended by the manufacturer (specificity, 0.993; sensitivity, 0.985; false-positive rate, 0.015). 相似文献
6.
Detection of rotavirus with a new polyclonal antibody enzyme immunoassay (Rotazyme II) and a commercial latex agglutination test (Rotalex): comparison with a monoclonal antibody enzyme immunoassay.
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G V Doern J E Herrmann P Henderson D Stobbs-Walro D M Perron N R Blacklow 《Journal of clinical microbiology》1986,23(2):226-229
A total of 176 human fecal specimens were examined for the presence of rotavirus by four different assays: a monoclonal antibody enzyme immunoassay; the original polyclonal antibody enzyme immunoassay marketed by Abbott Laboratories, North Chicago, Ill. (Rotazyme I); a modification of this assay which is now commercially available (Rotazyme II); and a latex agglutination test (Rotalex) recently introduced by Medical Technology Corp., Somerset, N.J. In addition, selected specimens were examined for the presence of rotavirus by electron microscopy, immune electron microscopy, and RNA gel electrophoresis. A total of 40 specimens were positive in the monoclonal antibody enzyme immunoassay, and 136 were negative. Using the results obtained with this procedure as the reference standard, we found the sensitivities of the Rotazyme I, Rotazyme II, and Rotalex tests to be 97.4, 100, and 81.6%, respectively. The specificities of these three procedures were 88.8, 83.9, and 100%, respectively. 相似文献
7.
Eighty-two stool specimens obtained from children with gastrointestinal disease were tested for the presence of antigen to rotavirus by latex agglutination (LA) (Virogen (VR), Meritec (MER), Wellcome (WEL), Slidex Rotatest (SRT), and enzyme-linked immunosorbent assays (Rotaclone [TRC], Rotazyme II [RTZ], Pathfinder [PTH]). Confirmatory testing was performed by isolation of rotavirus from stool specimens with the use of a shell vial centrifugation, antigen-detection tissue culture amplification method. The sensitivities and negative predictive values of VR, MER, WEL, SRT, TRC, RTZ, and PTH tests were 85, 89, 95, 91, 98, and 100%, respectively. Each test demonstrated 100% specificity and positive predictive values except the SRT, which attained a specificity of 95%. The WEL LA test may be used as a preliminary rapid screening assay following a stat request. The Kallestad PTH ELISA, however, was determined to be the rotavirus antigen detection kit of choice for routine laboratory diagnostic testing. 相似文献
8.
Direct appraisal of latex agglutination testing, a convenient alternative to enzyme immunoassay for the detection of rotavirus in childhood gastroenteritis, by comparison of two enzyme immunoassays and two latex tests.
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During February and March 1984, 207 fecal samples from infants and children with gastroenteritis were tested for rotavirus with four techniques: two enzyme immunoassays (Rotazyme; Abbott Laboratories, North Chicago, Ill., and Enzygnost-Rotavirus; Calbiochem-Behring, La Jolla, Calif.) and two latex agglutination tests (Rotalex; Orion Research, Inc., Cambridge, Mass., and Slidex Rota-Kit; Biomérieux). All stool samples were also tested for yeasts and bacterial pathogens. Electron microscopy was used to investigate discrepant results. We found 47% positive samples with Enzygnost-Rotavirus, 38% with Rotazyme, 37% with Slidex Rota-Kit, and 34% with Rotalex. No specimen was found positive by Rotazyme only or Slidex Rota-Kit only. On the contrary, 12 samples which were positive with Enzygnost-Rotavirus only and 3 which were positive with Rotalex only were not confirmed as positive by electron microscopy. Both enzyme immunoassays gave 6% equivocal results; Slidex Rota-Kit gave significantly fewer equivocal results than did Rotalex: 2.9% versus 9.7% (P less than 0.01). The sensitivity and specificity of latex tests compared favorably with that of enzyme immunoassays. Latex agglutination tests can be performed by unskilled personnel and are rapid and relatively cheap. They appear to be very suitable for routine laboratory work and may prove useful for large-scale screening in developing countries. 相似文献
9.
Rapid diagnosis of rotavirus gastroenteritis by a commercial latex agglutination test. 总被引:1,自引:6,他引:1
The Rotalex test, a commercial latex agglutination test for rotavirus, was compared with direct electron microscopy (EM) and the Rotazyme test I, a commercial enzyme immunoassay, for detection of rotavirus in stools of children and neonates. For initial stool specimens from 265 children (less than 3 years old) with diarrhea, the Rotalex test had a sensitivity of 81.7% and specificity of 99.5% compared with EM results. Positive and negative predictive values were 98 and 94.9%, respectively. The Rotalex test was slightly more sensitive and specific than the Rotazyme test. When daily stool specimens from patients with rotavirus gastroenteritis were examined, the sensitivity of the Rotalex test varied depending on the time of stool collection relative to the onset of symptoms. Sensitivity was 100 (20/20), 96 (23/24), and 54% (7/13) during 1 to 4, 5 to 7, and 8 to 18 days, respectively, after the onset of symptoms. The sensitivity of the Rotazyme test varied similarly with days from onset. We also examined 214 EM-negative stool specimens from asymptomatic newborns. False positivity by the Rotalex test was only 3.3% (7/214) compared with 4.2% (9/215) for the Rotazyme test. The Rotalex test was as sensitive and specific as EM for detection of rotavirus during the acute stage of illness and much faster and cheaper than EM or the Rotazyme test. The test appears to be suitable for routine use in small hospitals, emergency wards, or even the physician's office for rapid diagnosis of rotavirus gastroenteritis. 相似文献
10.
Regagnon C Chambon M Archimbaud C Charbonné F Demeocq F Labbé A Aumaître O Ughetto S Peigue-Lafeuille H Henquell C 《Pathologie-biologie》2006,54(6):343-346
The ability of two commercially available diagnosis rapid assays in detecting rotavirus antigen was compared in a prospective study conducted from September 2002 to May 2003. Five hundred and twelve faecal specimens were studied by IDEIA Rotavirus enzyme immunoassay test (EIA) and Diarlex MB immunochromatographic test (ICG). Specimens giving discrepant results were examined by electron microscopy (EM) and clinical data reconsidered. Out of 512 stool specimens, 155 (30.3%) were positive and 332 (64.8%) negative with the two assays. Discrepant results were obtained for 25 (4.88%) specimens (24 children, 1 adult), with EIA giving more positive results. The retrospective examination by EM, possible for fifteen stools on the 25 that gave discrepant results, confirmed the presence of rotavirus in 7/14 stools which were positive only by EIA and in the stool specimen that was found positive only by ICG. The 25 clinical observations re-examination showed the presence of GEA signs in all cases. The statistical analysis shows an excellent concordance between the EIA and the ICG tests (kappa = 0.89, IC(95%) = [0.85-0.93]) in spite of the underestimation of ICG test in comparison with EIA test (P < 0.0001). 相似文献
11.
Comparative efficiency of commercial immunoassays for the diagnosis of rotavirus gastroenteritis during the course of infection.
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We evaluated the performance characteristics of three commercially available immunoassays for the detection of rotavirus antigens in stool samples obtained from infants during the course of rotavirus gastroenteritis. Two of the assays, Bio-EnzaBead (Litton Bionetics, Charleston, S.C.) and Rotazyme (Abbott Laboratories, North Chicago, Ill.), are enzyme immunoassays, while the third, Rotalex (Medical Technology Corporation, Somerset, N.J.), is a latex agglutination assay. We tested a total of 122 samples obtained from 26 children with gastroenteritis; 56 samples, obtained from 21 children, were found to contain rotavirus antigen by a reference microplate enzyme immunoassay. Rotavirus antigen was found by the Bio-EnzaBead, Rotazyme, and Rotalex assays in 53, 42, and 29 samples, respectively. The true positivity of samples which were positive by the reference microplate assay but negative by the other assays was confirmed by a specific neutralization assay or by the visualization of bands of double-stranded RNA by polyacrylamide gel electrophoresis or both. No false-positive assay results were noted with any of the commercial assays. The sensitivity of the assays was determined to a great extent by the time after the onset of illness at which the specimen was collected. Thus, the sensitivity of commercial assays with specimens collected early in the course of illness did not differ significantly from that of the microplate assay. However, significantly lower degrees of sensitivity were noted later in the course of illness. Results of our studies indicate that all three commercial assays can accurately detect rotavirus in stools from children with rotavirus gastroenteritis. However, the choice of assay systems for use in the clinical laboratory will depend on the conditions in which stool specimens are collected and tested in the laboratory. 相似文献
12.
Radioimmunoassay and enzyme immunoassay (EIA) are generally recommended for routine diagnosis of rotavirus infection in childhood gastroenteritis. Expensive and delicate, these techniques are ill-suited for processing a small number of samples. Recently, Latex agglutination tests have been introduced than can be performed by non specialized hospital laboratories. However, some questions have been raised as to the sensitivity and specificity of these tests. We have sought a direct appraisal of latex agglutination testing by comparing two enzyme immunoassays and two latex tests (Rotazyme, Enzygnost-Rotavirus, Rotalex, Slidex Rota-kit). Three comparative studies that involved 1217 stool samples from children with gastroenteritis were carried out. Specificity, sensitivity, of latex tests compared favorably with the more sophisticated EIA, they represent a very convenient alternative for routine laboratory use provided latex tests with low rate of non-specific agglutinations are chosen. 相似文献
13.
Peroxidase-labeled monoclonal antibody against rotavirus group-specific antigen (inner capsid) was used for the detection of rotavirus by immunoperoxidase staining (IPS) in trypsin-free MA104 cells within 18 h post-inoculation with clinical specimens. One hundred and twenty-one fecal samples from children with acute gastroenteritis were evaluated by IPS, conventional virus isolation in cell culture and a commercially available group A-antigen ELISA (Rotazyme II, Abbott Laboratories). Fifty-eight (47.9%) stool samples were found positive by IPS. In contrast, rotavirus was isolated from only 4 (3.3%) fecal specimens by conventional cell culture (i.e. demonstration of a cytopathogenic effect). A total of 93 (76.9%) samples were positive by ELISA. IPS permits rapid detection of rotavirus infections and detects shedding of infectious virus. The method should be useful for the investigation of nosocomial spread of rotavirus infection in hospitals, contamination of environmental surfaces and desinfectants. 相似文献
14.
Evaluation of a latex agglutination kit (Virogen Rotatest) for detection of bovine rotavirus in fecal samples 总被引:1,自引:0,他引:1
Al-Yousif Y Anderson J Chard-Bergstrom C Bustamante A Muenzenberger M Austin K Kapil S 《Clinical and diagnostic laboratory immunology》2001,8(3):496-498
The performance of the Virogen Rotatest latex agglutination test (LAT) was evaluated for detection of bovine rotavirus antigen. Sixty-three fecal samples from diarrheic calves were collected from November 1999 to May 2000 and screened by LAT, the Rotazyme II enzyme-linked immunosorbent assay (ELISA), and virus isolation (VI) followed by an anti-rotavirus fluorescent-antibody (FA) test to detect the presence of group A rotavirus antigen. Of the 63 samples screened by VI-FA, 33 (58%) tested positive for rotavirus antigen. When the results from the LAT were compared to those from VI-FA, the "gold standard" for detection of bovine rotavirus in fecal samples, the sensitivity and specificity were found to be 87.8 and 73.3%, respectively. Latex agglutination compared with ELISA (the reference method) showed 100% sensitivity and 96.3% specificity, and when ELISA was compared with VI, the sensitivity was 84.8% and the specificity was 73.3%. Latex agglutination is easy to perform in a short time and does not require expensive equipment or skilled personnel, and the reagents have long shelf lives. These factors make the LAT suitable and highly efficient for use in a clinical laboratory as a rapid screening test for bovine rotavirus. 相似文献
15.
A new immune complex dot assay (ICDA) using immune gold/silver staining is described for the sensitive and rapid detection of rotavirus in cell culture and stool specimens. The method involves spotting preformed antigen-antibody complexes onto nitrocellulose paper, followed by incubation with colloidal gold-labelled secondary antibody and silver enhancement. ICDA was sensitive and specific and detected rotavirus antigens over a wide range of concentrations. It was more sensitive than a conventional immunodot assay (CIDA) and two commercial enzyme immunoassays (EIA) based on testing serial dilutions of a positive stool specimen. Of 26 stool specimens tested ICDA detected rotavirus antigen in 17; 14 were positive by Pathfinder Rotavirus EIA, 16 by Testpack Rotavirus EIA, and direct electron microscopy (DEM) detected only 12. The ICDA offers improved sensitivity over commercial EIAs and DEM. 相似文献
16.
Haemorrhagic colitis and haemolytic-uraemic syndrome: false positive reaction with a rotavirus latex agglutination test.
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A stool sample from a child with haemorrhagic colitis and haemolytic-uraemic syndrome gave a positive reaction with the RotaScreen latex agglutination test in the absence of other evidence of rotavirus infection. When this test is performed on bloody specimens, positive reactions should be interpreted with caution and confirmed by other means. 相似文献
17.
Comparison of three enzyme immunoassays to tissue culture for the diagnosis of rotavirus gastroenteritis in infants and young children. 总被引:1,自引:0,他引:1
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Three enzyme immunoassays (EIAs), Rotazyme II, IDL, and Pathfinder, were evaluated for rotavirus detection in stool and rectal swab specimens from children with symptomatic gastroenteritis and compared with virus isolation in primary African green monkey kidney cells. Of 125 specimens tested, 49 were rotavirus positive by tissue culture isolation; of these 49, 40 were positive by Rotazyme II, 43 were positive by IDL, and 46 were positive by Pathfinder EIAs. As compared with tissue culture isolation, the Rotazyme II, IDL, and Pathfinder EIAs had sensitivities of 82, 88, and 94%, specificities of 90, 99, and 95%, and overall agreements of 86, 94, and 94%, respectively. 相似文献
18.
Evaluation of seven immunoassays for detection of rotavirus in pediatric stool samples 总被引:1,自引:7,他引:1
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The performance of seven commercially manufactured rotavirus assays was evaluated with 144 pediatric stool specimens and compared with electron microscopy (EM) findings. The four enzyme-linked immunosorbent assays used were Rotazyme II, Pathfinder, IDL rotavirus immunoassay, and Enzygnost (Behring) rotavirus assay. The three latex tests were Meritec rotavirus detection test, Virogen Rotatest, and Bartels rotavirus latex test. Test outcomes were compared with EM on the basis of sensitivity, specificity, positive-negative predictive value, and the kappa statistic. Relative to EM, Meritec had the highest specificity (97%), followed by Virogen (95%), IDL (91%), Pathfinder (85%), Behring (81%), Bartels (72%), and Rotazyme (71%). The sensitivities were as follows: Rotazyme (92%), Pathfinder (89%), Bartels (86%), Virogen (86%), Behring (82%), Meritec (71%), and IDL (75%). Patient age and sex did not influence test results. Owing to the absence of a true standard, the tests were also compared with each other on the basis of the kappa statistic, the frequency of positive test results, and the frequency of samples in which a test differed from all other tests. Using these measures, the assays could be classified into three groups with progressively decreasing utility: group 1 (Virogen, Meritec, IDL, and EM), group 2 (Pathfinder and Behring), and group 3 (Rotazyme and Bartels). Laboratory criteria were also compared. Latex tests were faster and required less equipment than enzyme-linked immunosorbent assays. The Virogen latex assay showed the best overall performance, which made it our choice for rapid and accurate rotavirus diagnosis. However, in children who have gastrointestinal symptoms with negative rotavirus test results, EM will be useful until such time as immunological tests for other enteric viruses are available. 相似文献
19.
Evaluation of a Latex Agglutination Kit (Virogen Rotatest) for Detection of Bovine Rotavirus in Fecal Samples
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Yousif Al-Yousif Joe Anderson Cindy Chard-Bergstrom Adrian Bustamante Margaret Muenzenberger Kimberly Austin Sanjay Kapil 《Clinical and Vaccine Immunology : CVI》2001,8(3):496-498
The performance of the Virogen Rotatest latex agglutination test (LAT) was evaluated for detection of bovine rotavirus antigen. Sixty-three fecal samples from diarrheic calves were collected from November 1999 to May 2000 and screened by LAT, the Rotazyme II enzyme-linked immunosorbent assay (ELISA), and virus isolation (VI) followed by an anti-rotavirus fluorescent-antibody (FA) test to detect the presence of group A rotavirus antigen. Of the 63 samples screened by VI-FA, 33 (58%) tested positive for rotavirus antigen. When the results from the LAT were compared to those from VI-FA, the “gold standard” for detection of bovine rotavirus in fecal samples, the sensitivity and specificity were found to be 87.8 and 73.3%, respectively. Latex agglutination compared with ELISA (the reference method) showed 100% sensitivity and 96.3% specificity, and when ELISA was compared with VI, the sensitivity was 84.8% and the specificity was 73.3%. Latex agglutination is easy to perform in a short time and does not require expensive equipment or skilled personnel, and the reagents have long shelf lives. These factors make the LAT suitable and highly efficient for use in a clinical laboratory as a rapid screening test for bovine rotavirus. 相似文献
20.
Ramani S Arumugam R Gopalarathinam N Mohanty I Mathew S Gladstone BP Jana AK Kuruvilla KA Kang G 《Journal of medical virology》2008,80(6):1099-1105
A distinct feature of neonatal rotavirus infection is the association of unusual strains that appear to be prevalent only in neonatal units and persist for long periods of time. The main aims of this study were to determine if rotavirus can be detected on environmental surfaces in the neonatal nursery and whether the infection occurs in mothers of infected and uninfected neonates. Thirty rotavirus positive neonates and an equal number of negative neonates were enrolled in this study. Stool samples from 15 mothers in each group and environmental swabs collected from the bed and surfaces around neonates were tested for rotavirus using single round and nested PCR for the VP6 gene. Rotavirus could be detected in environmental swabs using single round PCR for VP6 gene in 40% of neonates positive for rotavirus antigen by enzyme immunoassay (EIA) and 33.3% of EIA negative neonates. The detection rate was almost 100% using the nested VP6 PCR. Rotavirus was detected in maternal samples only if the nested VP6 PCR was used, with no significant difference between rates of rotavirus detection in maternal fecal samples of infected and uninfected neonates (p-0.4). Sequence analysis of nested VP6 amplicons from two environmental swabs revealed them to be closest in identity to G10P[11], the most common genotype causing infections in neonates in this setting. Interestingly, sequences of amplicons from maternal stool samples did not cluster with G10P[11] or other VP6 subgroup I strains but showed clustering with human strains of VP6 subgroup II. 相似文献